LOCUS       sequence-479915-        5166 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express TREM2-SmBiT Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5166)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5166)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: TREM2-SmBiT Vector
FEATURES             Location/Qualifiers
     source          1..5166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2055..2176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2066..2085)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2291..2324)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2334..2381
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2413..3207
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2467..2486)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3077..3096
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3424..3969
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3870..3889
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4086..4103
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4253..4301
                     /note="synthetic polyadenylation signal"
     CDS             complement(4325..4723)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4720..4729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4754..5111)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4768..4903
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4920..5111
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5068..5088
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgatcgccat ggagcctctc
     1201 cggctgctca tcttactctt tgtcacagag ctgtccggag cccacaacac cacagtgttc
     1261 cagggcgtgg cgggccagtc cctgcaggtg tcttgcccct atgactccat gaagcactgg
     1321 gggaggcgca aggcctggtg ccgccagctg ggagagaagg gcccatgcca gcgtgtggtc
     1381 agcacgcaca acttgtggct gctgtccttc ctgaggaggt ggaatgggag cacagccatc
     1441 acagacgata ccctgggtgg cactctcacc attacgctgc ggaatctaca accccatgat
     1501 gcgggtctct accagtgcca gagcctccat ggcagtgagg ctgacaccct caggaaggtc
     1561 ctggtggagg tgctggcaga ccccctggat caccgggatg ctggagatct ctggttcccc
     1621 ggggagtctg agagcttcga ggatgcccat gtggagcaca gcatctccag gagcctcttg
     1681 gaaggagaaa tccccttccc acccacttcc atccttctcc tcctggcctg catctttctc
     1741 atcaagattc tagcagccag cgccctctgg gctgcagcct ggcatggaca gaagccaggg
     1801 acacatccac ccagtgaact ggactgtggc catgacccag ggtatcagct ccaaactctg
     1861 ccagggctga gagacacggt ttctcaaggc agttcaggtg gtggcgggag cggaggtgga
     1921 ggctcgagcg gtgtgaccgg ctaccggctg ttcgaggaga ttctgtaagg ccgcgactct
     1981 agagtcgacc tgcaggcatg caagctgatt tggctgctgc caccgctgag caaggccgct
     2041 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2101 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2161 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2221 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcttaattaa
     2281 gaattctaat ataacttcgt atagcataca ttatacgaag ttatggatcc actgttgaca
     2341 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc
     2401 aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     2461 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     2521 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     2581 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     2641 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     2701 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     2761 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     2821 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     2881 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     2941 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3001 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3061 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3121 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3181 tatcgccttc ttgacgagtt cttctgattc gaaatgaccg accaagcgac gcccaaccgg
     3241 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3301 agaacatgtc aaacatgaga attacaactt atatcgtatg gggctgactt caggtgctac
     3361 atttgaagag ataaattgca ctgaaatcta gaaatatttt atctgattaa taagatgatc
     3421 ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt
     3481 gcagggcggt ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct
     3541 tggaggagcg cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt
     3601 caagactaac tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc
     3661 tttccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg
     3721 ggggttcgtg catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc
     3781 gtggaatgag acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga
     3841 acaggagagc gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3901 gggtttcgcc accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc
     3961 ctatggaaaa acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat
     4021 cttccaggaa atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg
     4081 agcgtagcga gtcagtgagc gaggaagcgg aatatatcct gtatcacata ttctgctgac
     4141 gcaccggtgc agcctttttt ctcctgccac atgaagcact tcactgacac cctcatcagt
     4201 gccaacatag taagccagta tacactccgc tagcgctgat gtccggaatc gtcacacaaa
     4261 aaaccaacac acagatgtaa tgaaaataaa gatattttat tgcggccatc gtgatggcta
     4321 gagtttagcc ctcccacaca taaccagagg gcagcaattc acgaatccca actgccgtcg
     4381 gctgtccatc actgtccttc actatggctt tgatcccagg atgcagatcg agaagcacct
     4441 gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc atttccgatc gcgacgatac
     4501 aagtcaggtt gccagctgcc gcagcagcag cagtgcccag caccacgagt tctgcacaag
     4561 gtcccccagt aaaatgatat acattgacac cagtgaagat gcggccgtcg ctagagagag
     4621 ctgcgctggc gacgctgtag tcttcagaga tggggatgct gttgattgta gccgttgctc
     4681 tttcaatgag ggtggattct tcttgagaca aaggcttggc catggtggcg ctagtgtcag
     4741 aagaatcgag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     4801 atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat
     4861 ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg
     4921 ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     4981 ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc
     5041 ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac agctggttct
     5101 ttccgcctca gaaggtacct aaccaagttc ctctttcaga ggttatttca ggccatggtg
     5161 ctgcgc
//
